Compositions and methods for the therapy of Inflammatory Bowel Disease

ABSTRACT

Compositions and methods for the therapy of Inflammatory Bowel Disease (IBD), including Celiac Disease, Crohn&#39;s Disease, and Ulcerative Colitis, are disclosed. Illustrative compositions comprise one or more anti-type 1 interferon antagonists, such as anti-type 1 interferon receptor antibody antagonists and fragments thereof, as well as polypeptides and small molecules that inhibit the interaction of type 1 interferon with its receptor (IFNAR).

CROSS REFERENCE TO RELATED APPLICATION

This application claims the benefit of the filing date of U.S. Ser. No.60/465,155, filed Apr. 23, 2003, the entire contents of which areincorporated herein by reference.

BACKGROUND OF THE INVENTION

1. Technical Field of the Invention

The present invention relates generally to the therapy of CeliacDisease, Crohn's Disease, and Ulcerative Colitis (collectively referredto as Inflammatory Bowel Disease, or IBD). The invention is morespecifically related to antagonists of type-1 interferon as well as totherapeutic methods employing such antagonists for the treatment of IBD.

2. Description of Related Art

Celiac Disease, Crohn's Disease, and Ulcerative Colitis (collectivelyreferred to as Inflammatory Bowel Disease, or IBD) are chronic,inflammatory diseases of the gastrointestinal tract. While the clinicalfeatures vary somewhat between these two disorders, both arecharacterized by abdominal pain, diarrhea (often bloody), a variablegroup of ‘extra-intestinal’ manifestations (such as arthritis, uveitis,skin changes, etc) and the accumulation of inflammatory cells within thesmall intestine and colon (observed in pathologic biopsy or surgicalspecimens).

IBD affects both children and adults, and has a bimodal age distribution(one peak around 20, and a second around 40). IBD is a chronic, lifelongdisease, and is often grouped with other so-called “autoimmune”disorders (e.g. rheumatoid arthritis, type I diabetes mellitus, multiplesclerosis, etc). IBD is found almost exclusively in the industrializedworld. The most recent data from the Mayo Clinic suggest an overallincidence of greater than 1 in 100,000 people in the United States, withprevalence data in some studies greater than 1 in 1000. There is a cleartrend towards an increasing incidence of IBD in the US and Europe,particularly Crohn's Disease. The basis for this increase is notpresently clear. As such, IBD represents the 2^(nd) most commonautoimmune disease in the United States (after rheumatoid arthritis).

Type 1 interferons have been detected in the gut of patients withInflammatory Bowel Disease. For example, interferon alpha was reportedto be overexpressed in the gut mucosa of patients with Celiac Disease, agluten-sensitive enteropathy, and in the lamina propria of Crohn'sDisease patients. Monteleone et al., Gut 48: 425-429 (2001); Fais etal., J. Interferon Res. 14: 235-238 (1994). The biological significanceof the type 1 interferons in the tissues from these disease patients hasnot been described. Type 1 interferons have not been described in thecirculation of individuals with Inflammatory Bowel Disease and it isunclear what role, if any, interferon alpha plays in the pathology ofthese diseases.

Type 1 interferons (i.e. interferons alpha and beta) are multifunctionalcytokines that play a critical role in a variety of immune responsesystems. Abnormal production of type 1 interferons is associated withseveral pathological conditions including transplant rejection andautoimmune diseases such as rheumatoid arthritis, systemic lupuserythematosus, and insulin dependent diabetes. The biological effects oftype 1 interferons are mediated through a single cell-surface receptor(IFNAR) that binds to all of the type 1 interferons but not to the type2 interferon, interferon-γ. The type 1 interferon receptor is expressedat varying levels on all nucleated cells in the body. It is composed oftwo polypeptide chains designated IFNAR1 and IFNAR2, that, together,constitute the high-affinity receptor capable of transducing anintracellular signal upon interferon binding.

A mouse monoclonal antibody, designated 64G12, directed against theIFNAR1 chain of the human type 1 interferon receptor, has been shown toblock the activity of type 1 interferons by interfering with the bindingof the cytokines to their receptor. (See, U.S. Pat. Nos. 5,889,151,5,886,153, 5,731,169, 5,861,258, and 5,919,453, and 6,475,983, as wellas U.S. Patent Application Publication No. 20020055492, each of which isincorporated by reference herein in its entirety). In primatetransplantation models, 64G12, given in conjunction with cyclosporine,has provided remarkable long-term efficacy in prevention of skinallograft rejection and graft-versus host disease. Benizri et al., J.Interferon Cytokine Res. 18: 273 (1998).

Treatment of IBD is varied. First line therapy typically includessalicylate derivatives (e.g., 5-ASA) given orally or rectally. Responserates in uncomplicated Crohn's Disease are approximately 40% (comparedto 20% for placebo). Corticosteroids remain a mainstay in the treatmentof patients with more “refractory” disease, despite the untowardside-effects. Newer treatment options include anti-metabolites (e.g.,methotrexate, 6-mercaptopurine) and immunomodulators (e.g., Remicade—achimeric human antibody directed at the TNFα receptor).

In spite of considerable research into therapies for these disorders,Celiac Disease, Crohn's disease and ulcerative cholitis remain difficultto treat effectively. Accordingly, there remains an unmet need in theart for improved methods for treating such Inflammatory Bowel Diseases.The present invention fulfills these and other related needs.

SUMMARY OF THE INVENTION

The present invention provides compositions and methods for thetreatment of Inflammatory Bowel Disease, including, for example CeliacDisease, Crohn's disease and ulcerative colitis. Compositions of thepresent invention comprise one or more type 1 interferon antagonist,such as, for example, anti-type 1 interferon antibodies, anti-IFNARantibodies, fragments of any of the aforementioned antibodies, proteinsand small molecules. Within some embodiments, antagonists according tothe present invention may be chimeric, primatized, humanized,de-immunized and/or human antibodies or receptor binding fragmentthereof. Within other embodiments, the present invention providestherapeutic methods comprising the step of administering to a patientafflicted with IBD, a therapeutically effective amount of a type 1interferon antagonist. Still further embodiments provide therapeuticmethods comprising the steps of (a) administering to a patient afflictedwith IBD, a tolerizing amount of a type 1 interferon antagonist and (b)administering to the patient a therapeutically effective amount of atype 1 interferon antagonist.

Thus, within certain embodiments of the present invention are providedantagonists that interfere with type 1 interferon ligand binding suchas, for example, soluble receptor chains (e.g. soluble IFNAR2). Otherrelated embodiments provide antibodies or antigen binding fragmentsthereof that selectively bind to one or more type 1 interferon or bindto the IFNAR receptor in such a way that they interfere with ligandbinding, such as, for example, by competitive, non-competitive oruncompetitive inhibition. Alternative embodiments provide antagoniststhat interfere with signal transduction by the IFNAR receptor. Stillfurther embodiments provide antagonists that antagonize the downstreameffects of type 1 interferons.

Suitable antibody antagonists for use in the therapeutic methods of thepresent invention include monoclonal antibodies such as, for example,non-human, chimeric, primatized, humanized, de-immunized and/or fullyhuman antibodies or antigen binding fragments thereof. Antibodyantagonists may further comprise one or more chemical modifications toincrease the antibody's, or antigen binding fragment thereof, half-lifein circulation such as, for example, crosslinking to polyethylene glycol(i.e. PEGylation).

Within certain preferred embodiments, the antagonist is an antibody thatbinds at or near the IFNAR1 antigenic epitope recognized by the murinemonoclonal antibody designated 64G12 and/or the engineered human variantdesignated CPI-1697. The 64G12 monoclonal antibody was deposited at theECACC (European Collection of Animal Cell Cultures Porton DownSalisbury, Wiltshire SP4 056, United Kingdom) on Feb. 26, 1992.

Further embodiments of the present invention further comprise one ormore additional therapeutic such as, for example, an immunosuppressive,an anti-inflammatory, a steroid, an immunomodulatory agent, a cytokine,and a TNF antagonist. Exemplary immunosuppressives include azathioprine,methotrexate, cyclosporine, FK506, rapamycin, and mycophenolate mofetil.Exemplary anti-inflammatories include 5-aminosalicylic acid,sulfasalazine and olsalazine. Exemplary steroids includecorticosteroids, glucocorticosteroids, prednisone, prednisolone,hydrocortisone, methylprednisolone, dexamethasone and ACTH. Exemplaryimmunomodulatory agents include PVAC, anti-CD40 ligand, anti-CD40,natalizumab (Antegren™), anti-VCAM1 and anti-ICAM1. Exemplary cytokinesinclude IL-10. Exemplary TNF antagonists include infliximab (Remicade®),etanercept (Enbrel®), adalimumab (Humira™), and CDP870.

By other embodiments of the present invention are provided methods forthe treatment of an Inflammatory Bowel Disease such as, for example,Celiac Disease, Crohn's Disease, and ulcerative colitis which methodscomprise the step of administering to a patient afflicted with anInflammatory Bowel Disease a therapeutically effective amount of a type1 interferon antagonist as disclosed herein above.

By the methods of the present invention, the antagonist may beadministered by any suitable route of delivery so as to ensureappropriate bioavailability. Thus, within certain embodiments, suitableroutes of administration may include intravenous bolus, intravenous slowbolus, or infusion. By other embodiments, administration of the type 1interferon antagonist may be achieved through subcutaneous,intramuscular, transdermal or intradermal injection. Alternativeembodiments provide that administration may be achieved through mucosaldelivery such as, for example, through inhalation, or throughnasopharyngeal or oral administration.

Within certain embodiments employing a protein antagonist, such as, forexample, an antibody and/or an antigen binding fragment thereof, theroute of administration may be subcutaneous, intramuscular and/orintravenous. Intravenous administration may be as a bolus injection, aslow bolus injection or as an infusion. Alternative embodiments providethat the protein antagonists may be delivered transdermally,intradermally, and mucosally.

Exemplary dosages may be between 0.1 and 50 mg/kg body weight,inclusive, more preferably between 0.5 and 10 mg/kg body weight,inclusive, and still more preferably between 2 and 5 mg/kg, inclusive.Within certain embodiments, multiple repeat doses may be administered.

Within embodiments of the present invention employing proteinantagonists, the dosing frequency may be in the range of once per day toonce per month, inclusive, more preferably, in the range of twice perweek to every two weeks, inclusive, and still more preferablyapproximately once per week. Alternatively, the antagonist may be dosedat approximately the in vivo half-life if provided adequate exposure.

Certain other embodiments of the present invention provide that theantagonist may be administered in combination with other therapeuticssuch as, for example, an immunosuppressive, an anti-inflammatory, asteroid, an immunomodulatory agent, a cytokine, and a TNF antagonistsuch as those identified herein above.

Still further embodiments of the present invention provide methods fortreating a patient suffering from an Inflammatory Bowel Disease whichmethods comprise the steps of (a) administering a tolerizing dose of aprotein-based type 1 interferon antagonist and (b) administering atherapeutically effective dose of said protein-based type 1 interferonantagonist. Within preferred embodiments of these methods, theinterferon antagonist may be an antibody against the type 1 interferonreceptor (IFNAR). Exemplary anti-type 1 interferon antibodies includechimeric, primatized, humanized, de-immunized and human antibodies.Certain preferred anti-IFNAR antibodies include those that bind toIFNAR1 such as, for example, the murine monoclonal antibody designated64G12 and/or the engineered human variant designated CPI-1697.

Preferred ranges for the tolerizing dose of the protein-based type 1interferon antagonist are between 10 mg/kg body weight to 50 mg/kg bodyweight, inclusive. More preferred ranges for the tolerizing dose arebetween 20 mg/kg body weight and 40 mg/kg body weight, inclusive. Stillmore preferred ranges for the tolerizing dose are between 20 and 25mg/kg body weight, inclusive.

Within these therapeutic regimens, the therapeutically effective dose ofanti-type 1 interferon is preferably administered in the range of 0.1 to10 mg/kg body weight, inclusive. More preferred therapeuticallyeffective doses are in the range of 0.2 to 5 mg/kg body weight,inclusive. Still more preferred therapeutically effective doses are inthe range of 0.5 to 2 mg/kg body weight, inclusive. Within alternativeembodiments, the subsequent therapeutic dose or doses may be in the sameor different formulation as the tolerizing dose and/or may beadministered by the same or different route as the tolerizing dose.Preferably the therapeutic doses are administered intravenously,intramuscularly, or subcutaneously.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A-1D and 2A-2D are graphs showing the effect of CPI-1697 on bodyweights of IBD-afflicted CTT. FIGS. 1A-1D show results for Phase Istudies. FIGS. 2A-2D show results for Phase II studies. Percent bodyweight change of each animal was calculated using its body weight on Day0, the dosing initiation day as the baseline. Percent individual andgroup mean body weight changes and group mean body weights of survivinganimals were plotted. Statistical analyses (one way ANOVA) wereperformed for all time points for the treated and control groups. **: AtWeek 55 of Phase I study, treated animals had statistically significantbody weight changes compared with the controls (p<0.01). Arrowsrepresent the dosing schedules.

FIGS. 3A-3D and 4A-4D are graphs showing the effect of CPI-1697 ondiarrhea scores of IBD-afflicted CTT. FIGS. 3A-3D show results for PhaseI studies. FIGS. 4A-4D show results for Phase II studies. Weekly averagediarrhea scores (average of five week days) of surviving animal wereplotted for the control and treated groups. Group mean weekly averagediarrhea scores and percent group mean weekly diarrhea score change withWeek-1, right before the dosing initiation as the baseline, were alsoplotted. Arrows represent the dosing schedules.

FIGS. 5A-5D and 6A-6D are graphs showing the effect of CPI-1697 onactivity scores of IBD-afflicted CTT. FIGS. 5A-5D show results for PhaseI studies. FIGS. 6A-6D show results for Phase II studies. Activityscores representing neutrophil infiltration (average of three biopsysamples) of surviving animal were plotted for the control and treatedgroups. Group mean activity scores and percent group mean weeklyactivity score change with Week-1 for Phase I and Week-2 for Phase II,before the dosing initiation as the baseline, were also plotted. Arrowsrepresent the dosing schedules.

FIGS. 7A-7D and 8A-8D are graphs showing the effect of CPI-1697 onchronicity scores of IBD-afflicted CTT. FIGS. 7A-7D show results forPhase I studies. FIGS. 8A-8D show results for Phase II studies.Chronicity scores representing extent of permanent changes to the colonmorphology, including loss of crypts and alterations in glandularstructures (average of three biopsy samples) of surviving animal wereplotted for the control and treated groups. Group mean activity scoresand percent group mean weekly chronicity score change with Week-1 forPhase I and Week-2 for Phase II, before the dosing initiation as thebaseline, were also plotted. Arrows represent the dosing schedules.

FIGS. 9A-9D and 10A-10D are graphs showing the effect of CPI-1697 onhyperplasia scores of IBD-afflicted CTT. FIGS. 9A-9D show results forPhase I studies. FIGS. 10A-10D show results for Phase II studies.Hyperplasia scores representing abnormal increase in mucosal tissuethickness, including cellular and interstitial tissue (average of threebiopsy samples) of surviving animal were plotted for the control andtreated groups. Group mean activity scores and percent group mean weeklyhyperplasia score change with Week-1 for Phase I and Week-2 for PhaseII, before the dosing initiation as the baseline, were also plotted.Arrows represent the dosing schedules.

FIGS. 11A-11B are graphs showing serum CPI-1697 drug levels ofindividual animals in Phase I (FIG. 11A) and Phase II (FIG. 11B)studies.

FIGS. 12A-12B are bar graphs showing relative PAHA response levels ofCPI-1697 treated animals in Phase I (FIG. 12A) and Phase II (FIG. 12B)studies.

FIG. 13 is a graph showing normalized IFNAR1 expression levels on Bcells of individual animals in the Phase II study. IFNAR1 expressionlevels on B cells of individual animals at various time-points werenormalized by the mean IFNAR1 levels of the control animals at eachtime-point with the assumption that IFNAR1 levels remained relativelystable on control animals.

FIGS. 14A-14B (SEQ ID NOS:1-2) show the amino acid sequences of theheavy chain (H3) (FIG. 14A) (SEQ ID NO: 1) and light chain (K1) (FIG.14B) (SEQ ID NO:2) of the humanized anti-IFNAR-1 antibody CPI-1697. TheCDRs are underlined.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is directed generally to compositions and theiruse in therapeutic methods for the treatment of Inflammatory BowelDisease (IBD), particularly Celiac disease, Crohn's disease, andulcerative colitis. As described further below, illustrativecompositions of the present invention include type I interferonantagonists, particularly anti-IFNAR antibodies, anti-type I interferonantibodies and/or antigen binding fragments thereof as well aspolypeptides and small molecules that function as type I interferonantagonists. Without wishing to be limited to any particular theory ofoperation, exemplary inventive antagonists may interfere and/or competewith ligand binding and/or with interferon-mediated signal transduction.Preferably, type I interferon antagonists of the present invention havea long in vivo half-life in circulation and, as a consequence thereof,are effective in achieving a prolonged therapeutic response. Methods forextending in vivo antibody half-lives include, for example, constructionof fusion proteins such as immunoglobulin Fc fusions or conjugation topolyethylene glycol (PEGylation).

The present invention further provides therapeutic methods of use whichmethods employ one or more type I interferon antagonist, as indicatedabove, in the treatment of IBD. Exemplary methods provide long-termresponse to the type I interferon antagonist. Additionally, providedherein are therapeutic methods of use wherein delivery of one or moretype I interferon antagonist is employed in a tolerizing regimen, thatprevents and/or minimizes an immune response to the therapeutic protein,which tolerizing regimen is followed by a therapeutic regimen.

The practice of the present invention will employ, unless indicatedspecifically to the contrary, conventional methods of virology,immunology, microbiology, molecular biology and recombinant DNAtechniques within the skill of the art, many of which are describedbelow for the purpose of illustration. Such techniques are explainedfully in the literature. See, e.g., Sambrook, et al. Molecular Cloning:A Laboratory Manual (2nd Edition, 1989); Maniatis et al. MolecularCloning: A Laboratory Manual (1982); DNA Cloning: A Practical Approach,vol. I & II (D. Glover, ed.); Oligonucleotide Synthesis (N. Gait, ed.,1984); Nucleic Acid Hybridization (B. Hames & S. Higgins, eds., 1985);Transcription and Translation (B. Hames & S. Higgins, eds., 1984);Animal Cell Culture (R. Freshney, ed., 1986); Perbal, A Practical Guideto Molecular Cloning (1984).

All publications, patents and patent applications cited herein, whethersupra or infra, are hereby incorporated by reference in their entirety.

As used in this specification and the appended claims, the singularforms “a,” “an” and “the” include plural references unless the contentclearly dictates otherwise.

Antibody Type 1 Interferon Antagonists

As noted above, the present invention is directed generally tocompositions comprising antagonists of type 1 interferons as well astherapeutic methods of use that employ such compositions for thetreatment of Inflammatory Bowel Disease (IBD), particularly Celiacdisease, Crohn's disease, and ulcerative colitis. Within certainembodiments of the present invention, type 1 interferon antagonistsinclude anti-IFNAR antibodies and/or fragments thereof that bind to atype 1 interferon receptor and thereby block the binding of its ligand(i.e. interferon alpha, interferon beta or interferon omega).Alternatively or additionally, type 1 interferon antagonists may beanti-type 1 interferon antibodies and/or fragments thereof that bind toa type 1 interferon (i.e. interferon alpha, interferon beta orinterferon omega) and thereby block its binding to its receptor (i.e.IFNAR). Antibody-mediated inhibition of ligand binding may occur throughcompetitive, non-competitive or uncompetitive inhibition. Alternatively,antibody-based antagonists may act by preventing intracellular signalingthrough the type 1 interferon receptor.

Thus, included within the scope of the present invention are chimeric,primatized, veneered, humanized, deimmunized and human anti-IFNAR andanti-type 1 interferon antibodies and/or antigen-binding fragmentsthereof. Thus, and as disclosed further herein, inventive antibodiesencompass portions, variants and/or derivatives of any of the foregoingantibodies.

An antibody, or antigen-binding fragment thereof, is said to“specifically bind,” “immunogically bind,” and/or is “immunologicallyreactive” to a type 1 interferon receptor if it reacts at a detectablelevel (within, for example, an ELISA assay) to IFNAR or to a type 1interferon, but not to a type 2 interferon receptor, interferon-γ or toany other protein.

“Immunological binding,” as used herein, generally refers to thenon-covalent interactions of the type that occurs between an antibody,or fragment thereof, and the type 1 interferon or receptor for which theantibody is specific. The strength, or affinity, of immunologicalbinding interactions can be expressed in terms of the dissociationconstant (K_(d)) of the interaction, wherein a smaller K_(d) representsa greater affinity. Immunological binding properties of selectedantibodies can be quantified using methods well known in the art. Onesuch method entails measuring the rates of antigen-binding site/antigencomplex formation and dissociation, wherein those rates depend on theconcentrations of the complex partners, the affinity of the interaction,and on geometric parameters that equally influence the rate in bothdirections. Thus, both the “on rate constant” (K_(on)) and the “off rateconstant” (K_(off)) can be determined by calculation of theconcentrations and the actual rates of association and dissociation. Theratio of K_(off)/K_(on) enables cancellation of all parameters notrelated to affinity, and is thus equal to the dissociation constantK_(d). See, generally, Davies et al., Annual Rev. Biochem. 59: 439-473(1990).

An “antigen-binding site,” or “binding portion” of an antibody refers tothe part of the immunoglobulin molecule that participates in antigenbinding. The antigen binding site is formed by amino acid residues ofthe N-terminal variable (“V”) regions of the heavy (“H”) and light (“L”)chains. Three highly divergent stretches within the V regions of theheavy and light chains are referred to as “hypervariable regions” whichare interposed between more conserved flanking stretches known as“framework regions,” or “FRs”. Thus the term “FR” refers to amino acidsequences which are naturally found between and adjacent tohypervariable regions in immunoglobulins. In an antibody molecule, thethree hypervariable regions of a light chain and the three hypervariableregions of a heavy chain are disposed relative to each other in threedimensional space to form an antigen-binding surface. Theantigen-binding surface is complementary to the three-dimensionalsurface of a bound antigen, and the three hypervariable regions of eachof the heavy and light chains are referred to as“complementarity-determining regions,” or “CDRs.”

Antibodies may be prepared by any of a variety of techniques known tothose of ordinary skill in the art. See, e.g., Harlow and Lane,Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory (1988).In general, antibodies can be produced by cell culture techniques,including the generation of monoclonal antibodies as described herein,or via transfection of antibody genes into suitable bacterial ormammalian cell hosts, in order to allow for the production ofrecombinant antibodies. In one technique, an immunogen comprising type 1interferon receptor or portion thereof is initially injected into any ofa wide variety of mammals (e.g., mice, rats, rabbits, sheep, hamsters,goats or transgenic mice with human antibody repertoires).Alternatively, the immunogen may comprise cells or cell extractscontaining receptor, extracellular domains of the receptor (natural orrecombinant). A superior immune response may be elicited if type 1interferon receptor is joined to a carrier protein, such as bovine serumalbumin or keyhole limpet hemocyanin (KLH). The immunogen is injectedinto the animal host, preferably according to a predetermined scheduleincorporating one or more booster immunizations, and the animals arebled periodically. Immunization may be carried out with one or moreadjuvants such as complete and incomplete Freund's adjuvant. Polyclonalantibodies specific for the type 1 interferon receptor may then bepurified from such antisera by, for example, affinity chromatographyusing type 1 interferon receptor immunogenic regions coupled to asuitable solid support.

Monoclonal antibodies specific for a type 1 interferon receptor may beprepared, for example, using the technique of Kohler and Milstein,Nature 256(5517): 495-7 (1975), and improvements thereto. Briefly, thesemethods involve the preparation of immortal cell lines capable ofproducing antibodies having the desired specificity (i.e., reactivitywith the ganglioside of interest). Such cell lines may be produced, forexample, from spleen cells obtained from an animal immunized asdescribed above. The spleen cells are then immortalized by, for example,fusion with a myeloma cell fusion partner, preferably one that issyngeneic with the immunized animal. A variety of fusion techniques maybe employed. For example, the spleen cells and myeloma cells may becombined with a nonionic detergent for a few minutes and then plated atlow density on a selective medium that supports the growth of hybridcells, but not myeloma cells. A preferred selection technique uses HAT(hypoxanthine, aminopterin, thymidine) selection. After a sufficienttime, usually about 1 to 2 weeks, colonies of hybrids are observed.Single colonies are selected and their culture supernatants tested forbinding activity against a type 1 interferon receptor. Hybridomas havinghigh reactivity and specificity are preferred.

Monoclonal antibodies may be isolated from the supernatants of growinghybridoma colonies. In addition, various techniques may be employed toenhance the yield, such as injection of the hybridoma cell line into theperitoneal cavity of a suitable vertebrate host, such as a mouse.Monoclonal antibodies may then be harvested from the ascites fluid orthe blood. Contaminants may be removed from the antibodies byconventional techniques, such as chromatography, gel filtration,precipitation, and extraction. Anti-type 1 interferon receptor may beused in the purification process in, for example, an affinitychromatography step.

A number of therapeutically useful molecules are known in the art whichcomprise antigen-binding sites that are capable of exhibitingimmunological binding properties of an antibody molecule. Theproteolytic enzyme papain preferentially cleaves IgG molecules to yieldseveral fragments, two of which (the “F_(ab)” fragments) each comprise acovalent heterodimer that includes an intact antigen-binding site. Theenzyme pepsin is able to cleave IgG molecules to provide severalfragments, including the “F_(ab′2)” fragment which comprises bothantigen-binding sites. An “F_(v)” fragment can be produced bypreferential proteolytic cleavage of an IgM, and on rare occasions IgGor IgA immunoglobulin molecule. F_(v), F_(ab) and F_(ab′2) fragmentsare, however, more commonly derived using recombinant techniques knownin the art. The F_(v) fragment includes a non-covalent V_(H)::V_(L)heterodimer including an antigen-binding site which retains much of theantigen recognition and binding capabilities of the native antibodymolecule. Inbar et al. Proc. Nat. Acad. Sci. USA 69: 2659-2662 (1972);Hochman et al. Biochem 15: 2706-2710 (1976); and Ehrlich et al. Biochem19: 4091-4096 (1980).

A single chain F_(v) (“sF_(v)”) antibody is a covalently linkedV_(H)::V_(L) heterodimer which is expressed from a gene fusion includingV_(H)- and V_(L)-encoding genes linked by a peptide-encoding linker.Huston et al., Proc. Nat. Acad. Sci. USA 85(16): 5879-5883 (1988). Anumber of methods have been described to discern chemical structures forconverting the naturally aggregated—but chemically separated—light andheavy anti-anti-type 1 interferon receptor antibody chains from anantibody V region into an sF_(v) molecule which will fold into a threedimensional structure substantially similar to the structure of anantigen-binding site. See, e.g., U.S. Pat. Nos. 5,091,513 and 5,132,405,to Huston et al.; and U.S. Pat. No. 4,946,778, to Ladner et al.

Each of the above-described molecules includes a heavy chain and a lightchain CDR set, respectively interposed between a heavy chain and a lightchain FR set which provide support to the CDRs and define the spatialrelationship of the CDRs relative to each other. As used herein, theterm “CDR set” refers to the three hypervariable regions of a heavy orlight chain V region. Proceeding from the N-terminus of a heavy or lightchain, these regions are denoted as “CDR1,” “CDR2,” and “CDR3”respectively. An antigen-binding site, therefore, includes six CDRs,comprising the CDR set from each of a heavy and a light chain V region.A type 1 interferon antagonist comprising a single CDR, (e.g., a CDR1,CDR2 or CDR3) is referred to herein as a “molecular recognition unit.”Crystallographic analysis of a number of antigen-antibody complexes hasdemonstrated that the amino acid residues of CDRs form extensive contactwith bound antigen, wherein the most extensive antigen contact is withthe heavy chain CDR3. Thus, the molecular recognition units areprimarily responsible for the specificity of an antigen-binding site.

As used herein, the term “FR set” refers to the four flanking amino acidsequences that frame the CDRs of a CDR set of a heavy or light chain Vregion. Some FR residues may contact bound antigen; however, FRs areprimarily responsible for folding the V region into the antigen-bindingsite, particularly the FR residues directly adjacent to the CDRs. WithinFRs, certain amino acid residues and certain structural features arevery highly conserved. In this regard, all V region sequences contain aninternal disulfide loop of around 90 amino acid residues. When the Vregions fold into a binding-site, the CDRs are displayed as projectingloop motifs which form an antigen-binding surface. It is generallyrecognized that there are conserved structural regions of FRs whichinfluence the folded shape of the CDR loops into certain “canonical”structures—regardless of the precise CDR amino acid sequence. Further,certain FR residues are known to participate in non-covalent interdomaincontacts which stabilize the interaction of the antibody heavy and lightchains.

A number of “humanized” antibody molecules comprising an antigen-bindingsite derived from a non-human immunoglobulin have been described,including antibodies having their associated non-human V-regions fusedto human constant domains (Winter and Milstein Nature 349: 293-299(1991); Lobuglio et al. Proc. Nat. Acad. Sci. USA 86: 4220-4224 (1989);Shaw et al. J. Immunol. 138: 4534-4538 (1987); and Brown et al. CancerRes. 47: 3577-3583 (1987)), non-human CDRs grafted into a humansupporting FR prior to fusion with an appropriate human antibodyconstant domain (Riechmann et al. Nature 332: 323-327 (1988); Verhoeyenet al. Science 239: 1534-1536 (1988); and Jones et al. Nature 321:522-525 (1986)), and rodent CDRs supported by recombinantly veneeredrodent FRs (European Patent Publication No. 519,596, published Dec. 23,1992). These “humanized” molecules are designed to minimize unwantedimmunological response toward non-human antihuman antibody moleculeswhich limits the duration and effectiveness of therapeutic applicationsof those moieties in human recipients.

As used herein, the terms “veneered FRs” and “recombinantly veneeredFRs” refer to the selective replacement of FR residues from, e.g., arodent heavy or light chain V region, with human FR residues in order toprovide a xenogeneic molecule comprising an antigen-binding site whichretains substantially all of the native FR folding structure. Veneeringtechniques are based on the understanding that the ligand bindingcharacteristics of an antigen-binding site are determined primarily bythe structure and relative disposition of the heavy and light chain CDRsets within the antigen-binding surface. Davies et al. Ann. Rev.Biochem. 59: 439-473 (1990). Thus, antigen binding specificity can bepreserved in a humanized antibody only wherein the CDR structures, theirinteraction with each other, and their interaction with the rest of theV region domains are carefully maintained. By using veneeringtechniques, exterior (e.g., solvent-accessible) FR residues which arereadily encountered by the immune system are selectively replaced withhuman residues to provide a hybrid molecule that comprises either aweakly immunogenic, or substantially non-immunogenic veneered surface.

The process of veneering makes use of the available sequence data forhuman antibody variable domains compiled by Kabat et al., in Sequencesof Proteins of Immunological Interest, 4th ed., (U.S. Dept. of Healthand Human Services, U.S. Government Printing Office, 1987), updates tothe Kabat database, and other accessible U.S. and foreign databases(both nucleic acid and protein). Solvent accessibilities of V regionamino acids can be deduced from the known three-dimensional structurefor human and murine antibody fragments. There are two general steps inveneering a murine antigen-binding site. Initially, the FRs of thevariable domains of an antibody molecule of interest are compared withcorresponding FR sequences of human variable domains obtained from theabove-identified sources. The most homologous human V regions are thencompared residue by residue to corresponding murine amino acids. Theresidues in the murine FR which differ from the human counterpart arereplaced by the residues present in the human moiety using recombinanttechniques well known in the art. Residue switching is only carried outwith moieties which are at least partially exposed (solvent accessible),and care is exercised in the replacement of amino acid residues whichmay have a significant effect on the tertiary structure of V regiondomains, such as proline, glycine and charged amino acids.

In this manner, the resultant “veneered” murine antigen-binding sitesare thus designed to retain the murine CDR residues, the residuessubstantially adjacent to the CDRs, the residues identified as buried ormostly buried (solvent inaccessible), the residues believed toparticipate in non-covalent (e.g., electrostatic and hydrophobic)contacts between heavy and light chain domains, and the residues fromconserved structural regions of the FRs which are believed to influencethe “canonical” tertiary structures of the CDR loops. These designcriteria are then used to prepare recombinant nucleotide sequences whichcombine the CDRs of both the heavy and light chain of a murineantigen-binding site into human-appearing FRs that can be used totransfect mammalian cells for the expression of recombinant humanantibodies which exhibit the antigen specificity of the murine antibodymolecule.

The present invention also contemplates that it may be desirable toreduce the in vivo immunogenicity or any of the antibodies prepared asoutlined above or by methods otherwise available in the art. Oneexemplary approach for achieving antibodies having reducedimmunogenicity is the DeImmunisation™ methodology provided by Biovation(Aberdeen, U.K.). By this methodology, human helper T-cell epitopes thatcomprise MHC class II binding sequences are identified and removed fromtherapeutic antibodies in order to minimize activation anddifferentiation of helper T-cells when the antibody is administered invivo.

A preferred antibody of the invention is a humanized antibody referredto herein as CPI-1697. This antibody is composed of a heavy chainreferred to as H3 and a light chain referred to as K1. The amino acidsequences of the H3 heavy chain and K1 light chain variable regions areshown in FIGS. 14A (SEQ ID NO:1) and 14B (SEQ ID NO:2). The H3 heavychain contains the CDR1, CDR2 and CDR3 sequences from the heavy chain ofthe murine anti-IFNAR-1 antibody 64G12, grafted onto a consensus humanimmunoglobulin heavy chain framework sequence, whereas the K1 lightchain contains the CDR1, CDR2 and CDR3 sequences from the light chain ofthe murine anti-IFNAR-1 antibody 64G12, grafted onto a consensus humanimmunoglobulin kappa light chain framework sequence. The CPI-1697antibody further includes a human IgG4 constant region.

Other antibody-based IFNAR-1 antagonists suitable for use in theinvention are described in detail in the co-owned U.S. patentapplication entitled “Humanized Antibodies to Interferon AlphaReceptor-1 (IFNAR-1)”, Ser. No. 60/465,058, filed on Apr. 23, 2003, theentire contents of which are expressly incorporated herein by reference.

Small Molecule and Polypeptide Type 1 Interferon Antagonists

In addition to antibody-based type 1 interferon antagonists, the presentinvention also contemplates type 1 interferon antagonists, andcompositions thereof, comprising one or more small molecules such as,for example, those small molecules that interfere with binding of a type1 interferon with its receptor (i.e. IFNAR).

In certain embodiments, combinatorial libraries of potential smallmolecule antagonists may be screened for an ability to bind to a type 1interferon or to the type 1 interferon receptor. Conventionally, newchemical entities with useful properties are generated by identifying achemical compound (called a “lead compound”) with some desirableproperty or activity, e.g., inhibiting activity, creating variants ofthe lead compound, and evaluating the property and activity of thosevariant compounds. Often, high throughput screening (HTS) methods areemployed for such an analysis.

In one preferred embodiment, high throughput screening methods involveproviding a library containing a large number of potential therapeuticcompounds (candidate compounds). Such “combinatorial chemical libraries”are then screened in one or more assays to identify those librarymembers (particular chemical species or subclasses) that display adesired characteristic activity. The compounds thus identified can serveas conventional “lead compounds” or can themselves be used as potentialor actual IBD therapeutics.

A combinatorial chemical library is a collection of diverse chemicalcompounds generated by either chemical synthesis or biological synthesisby combining a number of chemical “building blocks” such as reagents.For example, a linear combinatorial chemical library, such as apolypeptide (e.g., mutein) library, is formed by combining a set ofchemical building blocks called amino acids in every possible way for agiven compound length (i.e., the number of amino acids in a polypeptidecompound). Millions of chemical compounds can be synthesized throughsuch combinatorial mixing of chemical building blocks. Gallop et al., J.Med. Chem. 37(9): 1233-1251 (1994).

Preparation and screening of combinatorial chemical libraries is wellknown to those of skill in the art. Such combinatorial chemicallibraries include, but are not limited to, peptide libraries (see, e.g.,U.S. Pat. No. 5,010,175, Furka, Pept. Prot. Res. 37: 487-493 (1991),Houghton et al., Nature, 354: 84-88 (1991)), peptoids (PCT PublicationNo. WO 91/19735), encoded peptides (PCT Publication No. WO 93/20242),random bio-oligomers (PCT Publication WO 92/00091), benzodiazepines(U.S. Pat. No. 5,288,514), diversomers such as hydantoins,benzodiazepines and dipeptides (Hobbs et al., Proc. Nat. Acad. Sci. USA90: 6909-6913 (1993)), vinylogous polypeptides (Hagihara et al., J.Amer. Chem. Soc. 114: 6568 (1992)), nonpeptidal peptidomimetics with aBeta-D-Glucose scaffolding (Hirschmann et al., J. Amer. Chem. Soc. 114:9217-9218 (1992)), analogous organic syntheses of small compoundlibraries (Chen et al., J. Amer. Chem. Soc. 116: 2661 (1994)),oligocarbamates (Cho, et al., Science 261: 1303 (1993)), and/or peptidylphosphonates (Campbell et al., J. Org. Chem. 59: 658 (1994)). See,generally, Gordon et al., J. Med. Chem. 37: 1385 (1994), nucleic acidlibraries (see, e.g., Strategene, Corp.), peptide nucleic acid libraries(see, e.g., U.S. Pat. No. 5,539,083), antibody libraries (see, e.g.,Vaughn et al., Nature Biotechnology 14(3: 309-314 (1996), andPCT/US96/10287), carbohydrate libraries (see, e.g., Liang et al.,Science 274: 1520-1522 (1996), and U.S. Pat. No. 5,593,853), and smallorganic molecule libraries (see, e.g., benzodiazepines, Baum, C&EN,January 18, page 33 (1993); isoprenoids, U.S. Pat. No. 5,569,588;thiazolidinones and metathiazanones, U.S. Pat. No. 5,549,974;pyrrolidines, U.S. Pat. Nos. 5,525,735 and 5,519,134; morpholinocompounds, U.S. Pat. No. 5,506,337; benzodiazepines, U.S. Pat. No.5,288,514; and the like).

Devices for the preparation of combinatorial libraries are commerciallyavailable (see, e.g., 357 MPS, 390 MPS, Advanced Chem Tech, LouisvilleKy., Symphony, Rainin, Woburn, Mass., 433A Applied Biosystems, FosterCity, Calif., 9050 Plus, Millipore, Bedford, Mass.).

A number of well known robotic systems have also been developed forsolution phase chemistries. These systems include automated workstationslike the automated synthesis apparatus developed by Takeda ChemicalIndustries, LTD. (Osaka, Japan) and many robotic systems utilizingrobotic arms (Zymate II, Zymark Corporation, Hopkinton, Mass.; Orca,Hewlett-Packard, Palo Alto, Calif.), which mimic the manual syntheticoperations performed by a chemist. The above devices, with appropriatemodification, are suitable for use with the present invention. Inaddition, numerous combinatorial libraries are themselves commerciallyavailable (see, e.g., ComGenex, Princeton, N.J., Asinex, Moscow, Ru,Tripos, Inc., St. Louis, Mo., ChemStar, Ltd, Moscow, RU, 3DPharmaceuticals, Exton, Pa., Martek Biosciences, Columbia, Md., etc.).

For detection of interferon-receptor interactions, assays that detectIFN-mediated signal transduction may be used such as IFN-mediatedinhibition of cell proliferation in cultured human tumor cell lines.Additionally, reporter gene assays may be used, for example, usingreporter genes expressed from an IFN-sensitive gene promoter. Lallemandet al., J. Leukocyte Biol. 60: 137-146 (1996). Suitable reporter genesinclude genes encoding luciferase and green fluorescent protein. In suchan assay, reporter gene expression is dependent on IFN activity and theIFN antagonist selectively inhibits IFN-stimulated gene expression.

High throughput assays for evaluating the presence, absence,quantification, or other properties of particular polypeptides are wellknown to those of skill in the art. Similarly, binding assays andreporter gene assays are similarly well known. Thus, e.g., U.S. Pat. No.5,559,410 discloses high throughput screening methods for proteins, U.S.Pat. No. 5,585,639 discloses high throughput screening methods fornucleic acid binding (i.e., in arrays), while U.S. Pat. Nos. 5,576,220and 5,541,061 disclose high throughput methods of screening forligand/antibody binding.

In addition, high throughput screening systems are commerciallyavailable (see, e.g., Zymark Corp., Hopkinton, Mass.; Air TechnicalIndustries, Mentor, Ohio; Beckman Instruments, Inc. Fullerton, Calif.;Precision Systems, Inc., Natick, Mass., etc.). These systems typicallyautomate procedures, including sample and reagent pipetting, liquiddispensing, timed incubations, and final readings of the microplate indetector(s) appropriate for the assay. These configurable systemsprovide high throughput and rapid start up as well as a high degree offlexibility and customization. The manufacturers of such systems providedetailed protocols for various high throughput systems. Thus, e.g.,Zymark Corp. provides technical bulletins describing screening systemsfor detecting the modulation of gene transcription, ligand binding, andthe like.

In one embodiment, modulators are proteins, often naturally occurringproteins or fragments of naturally occurring proteins. Thus, e.g.,cellular extracts containing proteins, or random or directed digests ofproteinaceous cellular extracts, may be used. In this way libraries ofproteins may be made for screening in the methods of the invention.Particularly preferred in this embodiment are libraries of bacterial,fungal, viral, and mammalian proteins, with the latter being preferred,and human proteins being especially preferred. Particularly useful testcompound will be directed to the class of proteins to which the targetbelongs, e.g., substrates for enzymes or ligands and receptors.

In a preferred embodiment, modulators are peptides of from about 5 toabout 30 amino acids, with from about 5 to about 20 amino acids beingpreferred, and from about 7 to about 15 being particularly preferred.The peptides may be digests of naturally occurring proteins as isoutlined above, random peptides, or “biased” random peptides. By“randomized” or grammatical equivalents herein is meant that the nucleicacid or peptide consists of essentially random sequences of nucleotidesand amino acids, respectively. Since these random peptides (or nucleicacids, discussed below) are often chemically synthesized, they mayincorporate any nucleotide or amino acid at any position. The syntheticprocess can be designed to generate randomized proteins or nucleicacids, to allow the formation of all or most of the possiblecombinations over the length of the sequence, thus forming a library ofrandomized candidate bioactive proteinaceous agents.

In one embodiment, the library is fully randomized, with no sequencepreferences or constants at any position. In a preferred embodiment, thelibrary is biased. That is, some positions within the sequence areeither held constant, or are selected from a limited number ofpossibilities. In a preferred embodiment, the nucleotides or amino acidresidues are randomized within a defined class, e.g., of hydrophobicamino acids, hydrophilic residues, sterically biased (either small orlarge) residues, towards the creation of nucleic acid binding domains,the creation of cysteines, for cross-linking, prolines for SH-3 domains,serines, threonines, tyrosines or histidines for phosphorylation sites,etc.

Compositions Comprising Type 1 Interferon Antagonists

In additional embodiments, the present invention concerns formulation ofone or more of the type 1 interferon antagonists disclosed herein inpharmaceutically-acceptable carriers for administration to a cell or ananimal, either alone, or in combination with one or more othermodalities of therapy. For example, depending on the particulartherapeutic regimen contemplated, compositions of the present inventionmay further comprise one or more additional therapeutic such as, forexample, an immunosuppressive, an anti-inflammatory, a steroid, animmunomodulatory agent, a cytokine, and a TNF antagonist. Exemplaryimmunosuppressives include azathioprine, methotrexate, cyclosporine,FK506, rapamycin, and mycophenolate mofetil. Exemplaryanti-inflammatories include 5-aminosalicylic acid, sulfasalazine andolsalazine. Exemplary steroids include corticosteroids,glucocorticosteroids, prednisone, prednisolone, hydrocortisone,methylprednisolone, dexamethasone, and ACTH. Exemplary immunomodulatoryagents include PVAC, anti-CD40 ligand, anti-CD40, natalizumab(Antegren™), anti-VCAM1, and anti-ICAM1. Exemplary cytokines includeIL-10. Exemplary TNF antagonists include infliximab (Remicade®),etanercept (Enbrel®), adalimumab (Humira™), and CDP870.

It will be understood that, if desired, a composition as disclosedherein may be administered in combination with other agents as well,such as, e.g., other proteins or polypeptides or variouspharmaceutically-active agents. In fact, there is virtually no limit toother components that may also be included, given that the additionalagents do not cause a significant adverse effect upon contact with thetarget cells or host tissues. The compositions may thus be deliveredalong with various other agents as required in the particular instance.Such compositions may be purified from host cells or other biologicalsources, or alternatively may be chemically synthesized as describedherein.

Therefore, in another aspect of the present invention, pharmaceuticalcompositions are provided comprising one or more of the antibodyproteins and/or small molecules described herein in combination with aphysiologically acceptable carrier.

It will be apparent that any of the pharmaceutical compositionsdescribed herein can contain pharmaceutically acceptable salts of thepolypeptides of the invention. Such salts can be prepared, for example,from pharmaceutically acceptable non-toxic bases, including organicbases (e.g., salts of primary, secondary and tertiary amines and basicamino acids) and inorganic bases (e.g., sodium, potassium, lithium,ammonium, calcium and magnesium salts).

While any suitable carrier known to those of ordinary skill in the artmay be employed in the compositions of this invention, the type ofcarrier will typically vary depending on the mode of administration.Compositions of the present invention may be formulated for anyappropriate manner of administration, including for example, oral,nasal, mucosal, intravenous, intraperitoneal, and intramuscularadministration.

Carriers for use within such pharmaceutical compositions arebiocompatible, and may also be biodegradable. In certain embodiments,the formulation preferably provides a relatively constant level ofactive component release. In other embodiments, however, a more rapidrate of release immediately upon administration may be desired. Theformulation of such compositions is well within the level of ordinaryskill in the art using known techniques. Illustrative carriers useful inthis regard include microparticles of poly(lactide-co-glycolide),polyacrylate, latex, starch, cellulose, dextran and the like. Otherillustrative delayed-release carriers include supramolecular biovectors,which comprise a non-liquid hydrophilic core (e.g., a cross-linkedpolysaccharide or oligosaccharide) and, optionally, an external layercomprising an amphiphilic compound, such as a phospholipid (see e.g.,U.S. Pat. No. 5,151,254 and PCT Publication Nos. WO 94/20078,WO/94/23701 and WO 96/06638). The amount of active compound containedwithin a sustained release formulation depends upon the site ofimplantation, the rate and expected duration of release and the natureof the condition to be treated or prevented.

In another illustrative embodiment, biodegradable microspheres (e.g.,polylactate polyglycolate) are employed as carriers for the compositionsof this invention. Suitable biodegradable microspheres are disclosed,for example, in U.S. Pat. Nos. 4,897,268; 5,075,109; 5,928,647;5,811,128; 5,820,883; 5,853,763; 5,814,344, 5,407,609 and 5,942,252.Modified hepatitis B core protein carrier systems such as described inPCT Publication No. WO/99 40934, and references cited therein, will alsobe useful for many applications. Another illustrative carrier/deliverysystem employs a carrier comprising particulate-protein complexes, suchas those described in U.S. Pat. No. 5,928,647, which are capable ofinducing a class I-restricted cytotoxic T lymphocyte responses in ahost.

The pharmaceutical compositions of the invention will often furthercomprise one or more buffers (e.g., neutral buffered saline or phosphatebuffered saline), carbohydrates (e.g., glucose, mannose, sucrose ordextrans), mannitol, proteins, polypeptides or amino acids such asglycine, antioxidants, bacteriostats, chelating agents such as EDTA orglutathione, adjuvants (e.g., aluminum hydroxide), solutes that renderthe formulation isotonic, hypotonic or weakly hypertonic with the bloodof a recipient, suspending agents, thickening agents and/orpreservatives. Alternatively, compositions of the present invention maybe formulated as a lyophilizate.

The pharmaceutical compositions described herein may be presented inunit-dose or multi-dose containers, such as sealed ampoules or vials.Such containers are typically sealed in such a way to preserve thesterility and stability of the formulation until use. In general,formulations may be stored as suspensions, solutions or emulsions inoily or aqueous vehicles. Alternatively, a pharmaceutical compositionmay be stored in a freeze-dried condition requiring only the addition ofa sterile liquid carrier immediately prior to use.

The development of suitable dosing and treatment regimens for using theparticular compositions described herein in a variety of treatmentregimens, including e.g., oral, intravenous, intranasal, andintramuscular administration and formulation, is well known in the art,some of which are briefly discussed below for general purposes ofillustration.

In certain applications, the pharmaceutical compositions disclosedherein may be delivered via oral administration to an animal. As such,these compositions may be formulated with an inert diluent or with anassimilable edible carrier, or they may be enclosed in hard- orsoft-shell gelatin capsule, or they may be compressed into tablets, orthey may be incorporated directly with the food of the diet.

The active compounds may even be incorporated with excipients and usedin the form of ingestible tablets, buccal tables, troches, capsules,elixirs, suspensions, syrups, wafers, and the like (see, for example,Mathiowitz et al., Nature 1997 Mar. 27; 386(6623): 410-4; Hwang et al.,Crit Rev Ther Drug Carrier Syst 1998; 15(3): 243-84; U.S. Pat. No.5,641,515; U.S. Pat. No. 5,580,579 and U.S. Pat. No. 5,792,451).Tablets, troches, pills, capsules and the like may also contain any of avariety of additional components, for example, a binder, such as gumtragacanth, acacia, cornstarch, or gelatin; excipients, such asdicalcium phosphate; a disintegrating agent, such as corn starch, potatostarch, alginic acid and the like; a lubricant, such as magnesiumstearate; and a sweetening agent, such as sucrose, lactose or saccharinmay be added or a flavoring agent, such as peppermint, oil ofwintergreen, or cherry flavoring. When the dosage unit form is acapsule, it may contain, in addition to materials of the above type, aliquid carrier. Various other materials may be present as coatings or tootherwise modify the physical form of the dosage unit. For instance,tablets, pills, or capsules may be coated with shellac, sugar, or both.Of course, any material used in preparing any dosage unit form should bepharmaceutically pure and substantially non-toxic in the amountsemployed. In addition, the active compounds may be incorporated intosustained-release preparation and formulations.

Typically, these formulations will contain at least about 0.1% of theactive compound or more, although the percentage of the activeingredient(s) may, of course, be varied and may conveniently be betweenabout 1 or 2% and about 60% or 70% or more of the weight or volume ofthe total formulation. Naturally, the amount of active compound(s) ineach therapeutically useful composition may be prepared is such a waythat a suitable dosage will be obtained in any given unit dose of thecompound. Factors such as solubility, bioavailability, biologicalhalf-life, route of administration, product shelf life, as well as otherpharmacological considerations will be contemplated by one skilled inthe art of preparing such pharmaceutical formulations, and as such, avariety of dosages and treatment regimens may be desirable.

For oral administration the compositions of the present invention mayalternatively be incorporated with one or more excipients in the form ofa mouthwash, dentifrice, buccal tablet, oral spray, or sublingualorally-administered formulation. Alternatively, the active ingredientmay be incorporated into an oral solution such as one containing sodiumborate, glycerin and potassium bicarbonate, or dispersed in adentifrice, or added in a therapeutically-effective amount to acomposition that may include water, binders, abrasives, flavoringagents, foaming agents, and humectants. Alternatively the compositionsmay be fashioned into a tablet or solution form that may be placed underthe tongue or otherwise dissolved in the mouth.

In certain circumstances it will be desirable to deliver thepharmaceutical compositions disclosed herein intravenously orintramuscularly. Such approaches are well known to the skilled artisan,some of which are further described, for example, in U.S. Pat. No.5,543,158; U.S. Pat. No. 5,641,515 and U.S. Pat. No. 5,399,363. Incertain embodiments, solutions of the active compounds as free base orpharmacologically acceptable salts may be prepared in water suitablymixed with a surfactant, such as hydroxypropylcellulose. Dispersions mayalso be prepared in glycerol, liquid polyethylene glycols, and mixturesthereof and in oils. Under ordinary conditions of storage and use, thesepreparations generally will contain a preservative to prevent the growthof microorganisms.

Illustrative pharmaceutical forms suitable for injectable use includesterile aqueous solutions or dispersions and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersions (for example, see U.S. Pat. No. 5,466,468). In all cases theform must be sterile and must be fluid to the extent that easysyringability exists. It must be stable under the conditions ofmanufacture and storage and must be preserved against the contaminatingaction of microorganisms, such as bacteria and fungi. The carrier can bea solvent or dispersion medium containing, for example, water, ethanol,polyol (e.g., glycerol, propylene glycol, and liquid polyethyleneglycol, and the like), suitable mixtures thereof, and/or vegetable oils.Proper fluidity may be maintained, for example, by the use of a coating,such as lecithin, by the maintenance of the required particle size inthe case of dispersion and/or by the use of surfactants. The preventionof the action of microorganisms can be facilitated by variousantibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In manycases, it will be preferable to include isotonic agents, for example,sugars or sodium chloride. Prolonged absorption of the injectablecompositions can be brought about by the use in the compositions ofagents delaying absorption, for example, aluminum monostearate andgelatin.

In one embodiment, the solution should be suitably buffered if necessaryand the liquid diluent first rendered isotonic with sufficient saline orglucose. These particular aqueous solutions are especially suitable forintravenous and intramuscular administration. In this connection, asterile aqueous medium that can be employed will be known to those ofskill in the art in light of the present disclosure. For example, onedosage may be dissolved in 1 ml of isotonic NaCl solution and eitheradded to 1000 ml of hypodermoclysis fluid or injected at the proposedsite of infusion, (see for example, “Remington's PharmaceuticalSciences” 15th Edition, pages 1035-1038 and 1570-1580). Some variationin dosage will necessarily occur depending on the condition of thesubject being treated. Moreover, for human administration, preparationswill of course preferably meet sterility, pyrogenicity, and the generalsafety and purity standards as required by FDA Office of Biologicsstandards.

In another embodiment of the invention, the compositions disclosedherein may be formulated in a neutral or salt form. Illustrativepharmaceutically-acceptable salts include the acid addition salts(formed with the free amino groups of the protein) and which are formedwith inorganic acids such as, for example, hydrochloric or phosphoricacids, or such organic acids as acetic, oxalic, tartaric, mandelic, andthe like. Salts formed with the free carboxyl groups can also be derivedfrom inorganic bases such as, for example, sodium, potassium, ammonium,calcium, or ferric hydroxides, and such organic bases as isopropylamine,trimethylamine, histidine, procaine and the like. Upon formulation,solutions will be administered in a manner compatible with the dosageformulation and in such amount as is therapeutically effective.

The carriers can further comprise any and all solvents, dispersionmedia, vehicles, coatings, diluents, antibacterial and antifungalagents, isotonic and absorption delaying agents, buffers, carriersolutions, suspensions, colloids, and the like. The use of such mediaand agents for pharmaceutical active substances is well known in theart. Except insofar as any conventional media or agent is incompatiblewith the active ingredient, its use in the therapeutic compositions iscontemplated. Supplementary active ingredients can also be incorporatedinto the compositions. The phrase “pharmaceutically-acceptable” refersto molecular entities and compositions that do not produce an allergicor similar untoward reaction when administered to a human.

In certain embodiments, the pharmaceutical compositions may be deliveredby intranasal sprays, inhalation, and/or other aerosol deliveryvehicles. Methods for delivering genes, nucleic acids, and peptidecompositions directly to the lungs via nasal aerosol sprays has beendescribed, e.g., in U.S. Pat. No. 5,756,353 and U.S. Pat. No. 5,804,212.Likewise, the delivery of drugs using intranasal microparticle resins(Takenaga et al., J Controlled Release 1998 Mar. 2; 52(1-2): 81-7) andlysophosphatidyl-glycerol compounds (U.S. Pat. No. 5,725,871) are alsowell-known in the pharmaceutical arts. Likewise, illustrativetransmucosal drug delivery in the form of a polytetrafluoroetheylenesupport matrix is described in U.S. Pat. No. 5,780,045.

In certain embodiments, liposomes, nanocapsules, microparticles, lipidparticles, vesicles, and the like, are used for the introduction of thecompositions of the present invention into suitable hostcells/organisms. In particular, the compositions of the presentinvention may be formulated for delivery either encapsulated in a lipidparticle, a liposome, a vesicle, a nanosphere, or a nanoparticle or thelike. Alternatively, compositions of the present invention can be bound,either covalently or non-covalently, to the surface of such carriervehicles.

The formation and use of liposome and liposome-like preparations aspotential drug carriers is generally known to those of skill in the art(see for example, Lasic, Trends Biotechnol 1998 July; 16(7): 307-21;Takakura, Nippon Rinsho 1998 March; 56(3): 691-5; Chandran et al.,Indian J Exp Biol. 1997 August; 35(8): 801-9; Margalit, Crit Rev TherDrug Carrier Syst. 1995; 12(2-3): 233-61; U.S. Pat. No. 5,567,434; U.S.Pat. No. 5,552,157; U.S. Pat. No. 5,565,213; U.S. Pat. No. 5,738,868 andU.S. Pat. No. 5,795,587, each specifically incorporated herein byreference in its entirety).

In certain embodiments, liposomes are formed from phospholipids that aredispersed in an aqueous medium and spontaneously form multilamellarconcentric bilayer vesicles (also termed multilamellar vesicles (MLVs)).

Alternatively, in other embodiments, the invention provides forpharmaceutically-acceptable nanocapsule formulations of the compositionsof the present invention. Nanocapsules can generally entrap compounds ina stable and reproducible way (see, for example, Quintanar-Guerrero etal., Drug Dev Ind Pharm. 1998 December; 24(12): 1113-28). To avoid sideeffects due to intracellular polymeric overloading, such ultrafineparticles (sized around 0.1 μm) may be designed using polymers able tobe degraded in vivo. Such particles can be made as described, forexample, by Couvreur et al., Crit Rev Ther Drug Carrier Syst. 1988;5(1): 1-20; zur Muhlen et al., Eur J Pharm Biopharm. 1998 March; 45(2):149-55; Zambaux et al. J Controlled Release. 1998 Jan. 2; 50(1-3):31-40; and U.S. Pat. No. 5,145,684.

Therapeutic Methods for the Treatment of Inflammatory Bowel Diseases

As indicated herein above, the present invention also providestherapeutic methods for the treatment of Inflammatory Bowel Disease suchas, for example, Celiac Disease, Crohn's Disease, and ulcerativecolitis, which methods comprise the step of administering to a patientafflicted with IBD, a therapeutically effective amount of a compositioncomprising a type 1 interferon antagonist such as, for example, one ofthe antibody-based type 1 interferon antagonists described herein above.The present invention also provides, within further embodiments,therapeutic methods for the treatment of Inflammatory Bowel Diseasewhich methods comprise the steps of (a) administering to a patientafflicted with IBD, a tolerizing amount of a type 1 interferonantagonist and (b) administering to the patient a therapeuticallyeffective amount of a type 1 interferon antagonist. Furthermore, it maybe desirable to administer one or more type 1 interferon antagonists incombination with other therapeutics such as, for example, animmunosuppressive, an anti-inflammatory, a steroid, an immunomodulatoryagent, a cytokine, and a TNF antagonist such as those identified hereinabove.

Routes and frequency of administration of the therapeutic compositionsdescribed herein, as well as dosage, will vary from individual toindividual, and may be readily established using standard techniques. Bythe methods of the present invention, the antagonist may be administeredby any suitable route of delivery so as to ensure appropriatebioavailability. Thus, within certain embodiments, suitable routes ofadministration may include intravenous bolus, intravenous slow bolus, orinfusion. By other embodiments, administration of the type 1 interferonantagonist may be achieved through subcutaneous, intramuscular,transdermal or intradermal injection. Alternative embodiments providethat administration may be achieved through mucosal delivery such as,for example, through inhalation (e.g., by aspiration), or throughnasopharyngeal or oral administration.

Within certain embodiments employing a protein antagonist, such as, forexample, an antibody and/or an antigen binding fragment thereof, theroute of administration may be subcutaneous, intramuscular and/orintravenous. Intravenous administration may be as a bolus injection, aslow bolus injection, or as an infusion. Alternative embodiments providethat the protein antagonists may be delivered transdermally,intradermally, and mucosally.

In general, an appropriate dosage and treatment regimen provides theactive compound(s) in an amount sufficient to provide therapeuticbenefit. Such a response can be monitored by establishing an improvedclinical outcome (e.g., reductions in abdominal pain; bloody diarrhea;‘extra-intestinal’ manifestations such as arthritis, uveitis, and skinchanges, etc.; and in the accumulation of inflammatory cells within thesmall intestine and colon).

Depending on the precise nature of the treatment regimen, appropriatedosages of the type 1 interferon antagonists disclosed herein may bebetween 0.1 and 50 mg/kg body weight, inclusive, more preferably between0.5 and 10 mg/kg body weight, inclusive, and still more preferablybetween 2 and 5 mg/kg body weight, inclusive. Within certainembodiments, multiple repeat doses may be administered.

Within embodiments of the present invention employing proteinantagonists, the dosing frequency may be in the range of once per day toonce per month, inclusive, more preferably, in the range of twice perweek to every two weeks, inclusive, and still more preferablyapproximately once per week.

Still further embodiments of the present invention provide methods fortreating a patient suffering from an Inflammatory Bowel Disease whichmethods comprise the steps of (a) administering a tolerizing dose of atype 1 interferon antagonist wherein the first type 1 interferonantagonist is a protein antagonist and (b) administering atherapeutically effective dose of said type 1 interferon antagonist.Within preferred embodiments of these methods, the interferon antagonistmay be an antibody against the type 1 interferon receptor (IFNAR).Exemplary anti-type 1 interferon antibodies include chimeric,primatized, humanized, de-immunized and human antibodies. Certainpreferred anti-IFNAR antibodies include those that bind to IFNAR 1 suchas, for example, the murine monoclonal antibody designated 64G12 and/orthe engineered human variant designated CPI-1697.

To achieve an initial tolerizing dose, anti-type 1 interferon antibodiesmay be immunogenic in humans and in non-human primates. The immuneresponse may be biologically significant and may impair the therapeuticefficacy of the antibody even if the antibody is partly or chieflycomprised of human immunoglobulin sequences such as, for example, in thecase of a chimeric, primatized, or humanized antibody. Within certainpreferred embodiments, an initial high dose of antibody is administeredsuch that a degree of immunological tolerance to the therapeuticantibody is established. The tolerizing dose is sufficient to prevent orreduce the induction of an IgG antibody response to repeatadministration of the anti-IFNAR antibody.

Preferred ranges for the tolerizing dose of the first type 1 interferonantagonist are between 10 mg/kg body weight to 50 mg/kg body weight,inclusive. More preferred ranges for the tolerizing dose are between 20and 40 mg/kg, inclusive. Still more preferred ranges for the tolerizingdose are between 20 and 25 mg/kg, inclusive.

Within these therapeutic regimens, the therapeutically effective dose ofanti-type 1 interferon is preferably administered in the range of 0.1 to10 mg/kg body weight, inclusive. More preferred second therapeuticallyeffective doses are in the range of 0.2 to 5 mg/kg body weight,inclusive. Still more preferred therapeutically effective doses are inthe range of 0.5 to 2 mg/kg, inclusive. Within alternative embodiments,the subsequent therapeutic dose or doses may be in the same or differentformulation as the tolerizing dose and/or may be administered by thesame or different route as the tolerizing dose. Preferably thetherapeutic doses are administered intravenously, intramuscularly, orsubcutaneously.

The following Example is offered by way of illustration and not by wayof limitation.

EXAMPLE Use of IFNAR-1 Antagonist in the Treatment of Infammatory BowelDisease

Idiopathic colitis in the cotton-top tamarin (CTT; Sanguinus oedipus), anew-world primate species, is recognized in the art as a model ofInflammatory Bowel Disease (IBD) in humans. Afflicted animals have asimilar pathophysiology to ulcerative colitis, with similar histologicalchanges observed to the colon, and a common sequela in humans and CTT iscolon cancer. Colitis in CTT is associated with morbidity and mortality,both in the colitis phase and due to the colon cancer. The colitis ischaracterized by repetitive outbreaks of symptoms, with remission.Disease periods generally last about 4 weeks, although there issignificant individual variability. The clinical symptoms includediarrhea with blood in feces, with a maldigestion/malabsorptionsyndrome. Histologically, the disease is characterized by infiltrationof neutrophils into the mucosal epithelium of the large intestine, withprogressive degenerative changes to the morphology of the intestinalcrypts, which enable diagnosis of stage of progression of the disease.While the cause is not known, dietary and infectious agents probablyhave a role and almost certainly lead to exacerbation of the condition.

An engineered humanized from of the mouse 64G12 antibody, CPI-1697,(IgG4k), has been developed, which binds IFNAR1 and competes with 64G12for binding to a similar epitope. CPI-1697 was used to treat idiopathiccolitis in CTT as follows. Experimentally naïve animals were selectedfrom a colony, on the basis of a history of colitis and exhibitingclinical symptoms including diarrhea and weight loss at the time ofentry into the study. Animals underwent colon biopsy, confirminginflammation of the colon within two weeks prior to initiation of theexperiment. All study animals had positive histological colitis scoresof 2, in a range of 0-4, (0=normal tissue) for activity, hyperplasia andchronicity, according to an established quantification scheme. Madara etal., Gastroenterology 88: 13-19 (1985)). Animals were prescreened forIFNAR-1 levels prior to inclusion in the study by flow cytometry, usingphycoerythrin-conjugated CPI-1697 on isolated peripheral bloodleucocytes. CPI-1697 was sterile-filtered and prepared at 20 mg/mL invehicle solution (Dulbecco's Na PBS (sterile, USP).

The experiment was conducted in two sequential phases. In the firstexperiment (Phase I), 5 animals with colitis were treated with aninitial dose of CPI-1697 at 20 mg/kg, given by slow i.v. infusion,followed by seven 10 mg/kg doses administered i.m. twice weekly for 4weeks. Five control animals were administered equivalent volumes ofvehicle solution (Dulbecco's Na PBS) according to the same dosingschedule. In the second experiment (Phase II), 6 animals were treatedwith CPI-1697 under the same initial i.v. dose (20 mg/kg) followed bytwice-weekly i.m. doses (10 mg/kg) for 8 weeks. 5 control animals wereadministered vehicle solution according to the same schedule. In bothphases, animals were monitored for body weight (twice weekly), diarrheaand periodically by histologic scoring of colon biopsy.

In Phase I, animals were assessed both during the time of treatment andthen followed for 6 weeks after the end of the treatment period, and for4 weeks after the end of treatment in Phase II. All animals were alsofollowed-up further on weights and colon histology at 51 weeks and 27weeks after the end of treatment for Phase I and II, respectively.Diarrhea was graded and scored visually at least 5 times per week basedon a standardized 0-5 scale, where 0 represented normal fecal stools,and 5 represented very watery diarrhea. Venous blood samples were takenat regular intervals for analysis of primate anti-human antibody immuneresponses (PAHA). Colon biopsies were taken at three sites (1, 3 and 6cm from the distal end of the colon) at intervals during the treatmentand follow-up period, and the tissue was fixed in formalin, sectionedand stained with hematoxylin and eosin for histologic evaluation.

The sections were scored by a veterinary histopathologist, who wasblinded to the treatment groups. The scoring system, from 0 (normal) to4 (severe), used 3 independent criteria as follows (Madara et al.,1985). The first parameter was “activity”—number of infiltratingneutrophils, the second was “chronicity”—extent of permanent changes tothe colon morphology, including loss of crypts and alterations inglandular structures, which characteristically slowly increase over theduration of the course of colitis, and the third was“hyperplasia’—abnormal increase in mucosal tissue thickness, includingcellular and interstitial tissue. A mean histology score was determinedfrom the three biopsy levels, for each of the 3 parameters assessed, ateach time point.

Body weights for Phase I and Phase II are shown in FIGS. 1A-1D and2A-2D, respectively. Diarrhea scores for Phase I and Phase II are shownin FIGS. 3A-3D and 4A-4D, respectively. “Activity” scores for Phase Iand Phase II are shown in FIGS. 5A-5D and 6A-6D, respectively.“Chronicity” scores for Phase I and Phase II are shown in FIGS. 7A-7Dand 8A-8D, respectively. “Hyperplasia” scores for Phase I and Phase IIare shown in FIGS. 9A-9D and 10A-10D, respectively.

One animal (#25285) in the control group of Phase I was euthanizedduring the treatment period for an increased inguinal hernia unrelatedto ulcerative colitis or treatment regime. No animal died in the treatedgroup of Phase I. For Phase II, both the control and treated groups hadone animal die during the treatment period. Of the surviving animals,the treated group had a bigger percentage body weight increase than thecontrol group in both Phase I and Phase II study throughout the studyperiod (FIGS. 1A-1D and 2A-2D). The body weight increase was mostprominent 4 to 6 weeks after the end of dosing period. Mostinterestingly, long-term follow-up on the animals until 51 weeks afterthe dosing period for Phase I study and 27 or 43 weeks for Phase IIstudy showed even bigger body weight percent increase in the treatedanimals compared with the control animals. In particular, Phase Itreated animals had statistically significant body weight percentincrease over the controls (p<0.01). Three animals (#4199, 12300, 52099)in the treated group showing good body weight increase were found out tobe under 20 month at the initiation of the study and the body weightincrease could be due to normal growth of young animals.

The effect of CPI-1697 treatment on diarrhea scores of CTT is summarizedin FIGS. 3A-3D and 4A-4D. The group mean weekly average diarrhea scoreshowed an improvement (decrease in score) in the treated group duringthe course of the study in Phase II study whereas the control groupshowed no improvement (FIGS. 4A-4D). The improvements in diarrhea scoresstarted right after onset of treatment and tended to be sustained afterthe cessation of treatment in Phase II study. For Phase I study, theimprovement in diarrhea scores was not obvious for the treatment (FIGS.3A-3D).

The effect of CPI-1697 treatment on “activity” scores representingneutrophil infiltration in colon of CTT is summarized in FIGS. 5A-5D and6A-6D. In Phase I study (FIGS. 5A-5D), both the control and treatedgroup had decreased neutrophil infiltration right after initiation ofstudy. The treated group had increased neutrophil infiltration over theperiod of Week 0 to Week 8 and substantially decreased neutrophilinfiltration from Week 8 to Week 10. In contrast, the control group hadslightly decreased neutrophil infiltration over Week 0 to Week 10.However, by Week 55, the treated group had slightly less neutrophilinfiltration than the control group. In Phase II study (FIGS. 6A-6D),the treated group had decreased neutrophil infiltration (group mean)over the 12 week study period and further reduction by Week 35 whereasthe control group showed no decrease.

The effect of CPI-1697 treatment on “chronicity” scores representingextent of permanent changes to the colon morphology, including loss ofcrypts and alterations in glandular structures of CTT is summarized inFIGS. 7A-7D and 8A-8D. In Phase I study (FIGS. 7A-7D), both the controland treated group had improved colon morphology right after initiationof treatment, however such improvement was not sustained. The treatedgroup did not have any additional beneficial effect on colon morphologyover the control group. In Phase II study (FIGS. 8A-8D), the treatedgroup had improved colon morphology two weeks after initiation ofdosing, exactly the opposite of the control group. Moreover, the treatedgroup had more improvement in colon morphology than the control grouplong term at Week 35 compared to Week 12.

The effect of CPI-1697 treatment on “hyperplasia” scores representingabnormal increase in mucosal tissue thickness, including cellular andinterstitial tissue of CTT is summarized in FIGS. 9A-9D and 10A-10D. InPhase I study (FIGS. 9A-9D), similar to colon morphology change, boththe control and treated group had decreased mucosal tissue thicknessright after initiation of treatment, however such improvement was notsustained. The treated group did not have any additional beneficialeffect on mucosal tissue thickness over the control group. In Phase IIstudy (FIGS. 10A-10D), the treated group had similar changes as thecontrol group in mucosal tissue thickness over the study period of 12weeks. However, long-term follow-up showed that the treated group haddecreased mucosal tissue thickness compared to the control group longterm at Week 35.

Serum samples from each animal were assayed by ELISA for levels of drug(CPI-1697 anti-IFNAR-1 antibody) by detecting human IgG4, and forprimate-anti-human-antibody (PAHA) responses. As shown in FIGS. 11A-11B,drug levels of about 50-270 ng/ml of plasma were maintained throughoutthe treatment phase and PAHA levels were low or undetectable in themajority of animals (FIGS. 12A-12B). Although all animals received thesame dose on a weight basis, certain animals were observed to havehigher circulating levels of CPI-1697, of up to 3 times higher. Thisrange of concentrations of CPI-1697 has been demonstrated previously tobe effective in blocking IFNAR-1 in vitro in primate studies. Twoanimals (#8494 and #52099) in Phase I developed detectable PAHAresponses and no detectable PAHA response was observed in any of theanimals in Phase II (FIGS. 12A-12B). These results suggest that thedosing regimen used, including a high initial dose of CPI-1697 did notlead to a significant immunological response to this protein (humanantibody). Those two animals also had low drug levels (FIGS. 11A-11B).

Levels of IFNAR1 on white blood cells were assayed by flow cytometry andthe normalized receptor levels of Phase II study were summarized in FIG.13. Receptor levels of Phase I study were not obtained as the FACSassays were not optimized then. Interferon receptor blocking wasachieved to various levels, although not complete, in the treatedanimals. Animals having higher levels of serum CPI-1697 tended to havebetter receptor blocking.

In summary, in both Phase I and Phase II studies, CPI-1697 treatmentgenerated weight increase benefit over control in colitis-afflictedtamarins. Phase II study with longer CPI-1697 treatment period had evenbetter effect with improvement in diarrhea scores and histopathologyscores including “activity”, “chronicity” and “hyperplasia”. In Phase IIstudy, there was a correlation between positive clinical response andhigher circulating levels of CPI-1697. There was also a positivecorrelation between lack of PAHA response and improved clinical scores.Such correlations were not found for Phase I study. These correlationswere not testable by statistical methods due to the small sample size inthe study. Overall these results indicate that treatment with ahumanized antibody to IFNAR-1 produces a clinical improvement in coliticCTT. The effect was chronic rather than acute and trended to besustained beyond the treatment phases, as shown by long-term follow-upstudy. Longer-term treatment in Phase II study produced stronger effectsthan shorter treatments. Increased exposure, both in time and plasmalevels, to CPI-1697 was associated with greater response.

From the foregoing it will be appreciated that, although specificembodiments of the invention have been described herein for purposes ofillustration, various modifications may be made without deviating fromthe spirit and scope of the invention. Accordingly, the invention is notlimited except as by the appended claims.

1. A composition for the treatment of an Inflammatory Bowel Disease,comprising: (a) a therapeutically effective amount of a type 1interferon antagonist, and (b) a pharmaceutically acceptable carrier. 2.The composition of claim 1 wherein said Inflammatory Bowel Disease isselected from the group consisting of Celiac Disease, Crohn's disease,or ulcerative colitis.
 3. The composition of claim 1 wherein said type 1interferon antagonist is selected from the group consisting of anantibody, a polypeptide, and a small molecule.
 4. The composition ofclaim 3 wherein said type 1 interferon antagonist further comprises achemical modification selected from the group consisting of conjugationto polyethylene glycol and fusion to an immunoglobulin F_(c) region. 5.The composition of claim 1 wherein said type 1 interferon antagonist isan antibody and said antibody is selected from the group consisting of amonoclonal anti-type 1 interferon receptor antibody and a monoclonalanti-type 1 interferon antibody.
 6. The composition of claim 5 whereinsaid antibody is selected from the group consisting of a non-humanantibody, a chimeric antibody, a humanized antibody, and a humanantibody.
 7. The composition of claim 6 wherein said non-human antibodyis 64G12.
 8. The composition of claim 6 wherein said human antibody isCPI-1697.
 9. The composition of claim 1 further comprising a therapeuticselected from the group consisting of an immunosuppressive, ananti-inflammatory, a steroid, an immunomodulatory agent, a cytokine, anda TNF antagonist.
 10. The composition of claim 9 wherein saidimmunosuppressive is selected from the group consisting of azathioprine,methotrexate, cyclosporine, FK506, rapamycin, and mycophenolate mofetil.11. The composition of claim 9 wherein said anti-inflammatory isselected from the group consisting of 5-aminosalicylic acid,sulfasalazine, and olsalazine.
 12. The composition of claim 9 whereinsaid steroid is selected from the group consisting of corticosteroids,glucocorticosteroids, prednisone, prednisolone, hydrocortisone,methylprednisolone, dexamethasone, and ACTH.
 13. The composition ofclaim 9 wherein said immunomodulatory agent is selected from the groupconsisting of PVAC, anti-CD40 ligand, anti-CD40, natalizumab,anti-VCAM1, and anti-ICAM1.
 14. The composition of claim 9 wherein saidcytokine is IL-10.
 15. The composition of claim 9 wherein said TNFantagonist is selected from the group consisting of infliximab,etanercept, adalimumab, and CDP870.
 16. A method for the treatment of apatient afflicted with an Inflammatory Bowel Disease comprising the stepof administering to said patient a therapeutically effective amount of atype 1 interferon antagonist.
 17. The method of claim 16 wherein saidInflammatory Bowel Disease is selected from the group consisting ofCeliac Disease, Crohn's disease, and ulcerative colitis.
 18. The methodof claim 16 wherein said type 1 interferon antagonist is administered bya route selected from the group consisting of intravenous bolus,intravenous slow bolus, and infusion.
 19. The method of claim 16 whereinsaid type 1 interferon antagonist is administered by a route selectedfrom the group consisting of subcutaneously, intramuscularly,transdermally, intradermally, and intravenously.
 20. The method of claim16 wherein said type 1 interferon antagonist is administered by a routeof mucosal delivery selected from the group consisting of inhalation,nasopharyngeally, and orally.
 21. The method of claim 16 wherein saidtype 1 interferon antagonist is administered in a dosage range ofbetween 0.1 mg/kg body weight and 50 mg/kg body weight, inclusive. 22.The method of claim 21 wherein said type 1 interferon antagonist isadministered in a dosage range of between 0.5 mg/kg body weight and 10mg/kg body weight, inclusive.
 23. The method of claim 22 wherein saidtype 1 interferon antagonist is administered in a dosage range ofbetween 2 mg/kg body weight and 5 mg/kg body weight, inclusive.
 24. Themethod of claim 16 wherein said type 1 interferon antagonist isadministered at a frequency between once per day and once per month,inclusive.
 25. The method of claim 24 wherein said type 1 interferonantagonist is administered at a frequency between twice per week andevery two weeks, inclusive.
 26. The method of claim 24 wherein said type1 interferon antagonist is administered at a frequency of approximatelyonce per week.
 27. The method of claim 16 further comprisingadministration of a second therapeutic selected from the groupconsisting of an immunosuppressive, an anti-inflammatory, a steroid, animmunomodulatory agent, a cytokine, and a TNF antagonist.
 28. A methodfor the treatment of a patient afflicted with an Inflammatory BowelDisease comprising the steps of (a) administering to said patient at afirst time point a tolerizing dose of a first type 1 interferonantagonist and (b) administering to said patient at a second time pointa therapeutically effective dose of a second type 1 interferonantagonist.
 29. The method of claim 28 wherein said first type 1interferon antagonist is an anti-IFNAR antibody.
 30. The method of claim28 wherein said second type 1 interferon antagonist is an anti-type 1interferon antibody.
 31. The method of claim 28 wherein said tolerizingdose of said first type 1 interferon antagonist is sufficient to preventor reduce the induction of an IgG antibody response to repeatadministration of the first type 1 interferon antagonist.
 32. The methodof claim 28 wherein said tolerizing dose of said first type 1 interferonantagonist is between 10 mg/kg body weight to 50 mg/kg body weight,inclusive.
 33. The method of claim 32 wherein said tolerizing dose ofsaid first type 1 interferon antagonist is between 20 mg/kg body weightand 40 mg/kg body weight, inclusive.
 34. The method of claim 33 whereinsaid tolerizing dose of said first type 1 interferon antagonist isbetween 20 mg/kg body weight and 25 mg/kg body weight, inclusive. 35.The method of claim 28 wherein said therapeutically effective dose ofsaid second type 1 interferon antagonist is administered in the range of0.1 mg/kg body weight to 10 mg/kg body weight, inclusive.
 36. The methodof claim 35 wherein said therapeutically effective dose of said secondtype 1 interferon antagonist is administered in the range of 0.2 mg/kgbody weight to 5 mg/kg body weight, inclusive.
 37. The method of claim36 wherein said therapeutically effective dose of said second type 1interferon antagonist is administered in the range of 0.5 mg/kg bodyweight to 2 mg/kg body weight, inclusive.